Thrombin receptor antagonists

The present application claims priority under 35 USC section 119(e) of U.S. Provisional application Ser. No. 60/373,072, filed Apr. 16, 2002, the complete text, claims and figures of which are incorporated by reference herein as if fully set forth.

The present invention relates to substituted tricyclic thrombin receptor antagonists, pharmaceutical compositions containing them and their use in the treatment of diseases associated with thrombosis, atherosclerosis, restenosis, hypertension, angina pectoris, arrhythmia, heart failure, cerebral ischemia, stroke, inflammatory disorders, neurodegenerative diseases and cancer. The invention also relates to the combination of novel compounds of the invention and other cardiovascular agents.

Thrombin is known to have a variety of activities in different cell types and thrombin receptors are known to be present in such cell types as human platelets, vascular smooth muscle cells, endothelial cells and fibroblasts. It is therefore possible that thrombin receptor antagonists, also known as protease activated receptor (PAR) antagonists will be useful in the treatment of thrombotic, inflammatory, atherosclerotic and fibroproliferative disorders, as well as other disorders in which thrombin and its receptor play a pathological role.

Thrombin receptor antagonists peptides have been identified based on structure-activity studies involving substitutions of amino acids on thrombin receptors. In Bernatowicz et al, J. Med. Chem ., vol. 39, pp. 4879–4887 (1996), tetra- and pentapeptides are disclosed as being potent thrombin receptor antagonists, for example N-trans-cinnamoyl-p-fluoroPhe-p-guanidinoPhe-Leu-Arg-NH 2 and N-trans-cinnamoyl-p-fluoroPhe-p-guanidinoPhe-Leu-Arg-Arg-NH 2 . Peptide thrombin receptor antagonists are also disclosed in WO 94/03479, published Feb. 17, 1994.

Substituted tricyclic thrombin receptor antagonists are disclosed in U.S. Pat. Nos. 6,063,847, 6,326,380 and U.S. Ser. No. 09/880,222 (WO 01/96330) and 10/271,715.

The present invention relates to thrombin receptor antagonists represented by the Formula I:

or a pharmaceutically acceptable salt or solvate thereof, wherein:

the single dotted line represents an optional single bond;

represents an optional double bond;

n is 0–2;

Q is

R 1 is independently selected from the group consisting of H, (C 1 –C 6 )alkyl, fluoro(C 1 –C 6 )alkyl-, difluoro(C 1 –C 6 )alkyl-, trifluoro-(C 1 –C 6 )alkyl-, (C 3 –C 6 )cycloalkyl, (C 2 –C 6 )alkenyl, hydroxy-(C 1 –C 6 )alkyl-, and amino(C 1 –C 6 )alkyl-;

R 2 is independently selected from the group consisting of H, (C 1 –C 6 )alkyl, fluoro(C 1 –C 6 )alkyl-, difluoro(C 1 –C 6 )alkyl-, trifluoro-(C 1 –C 6 )alkyl-, (C 3 –C 6 )cycloalkyl, (C 2 –C 6 )alkenyl, hydroxy-(C 1 –C 6 )alkyl-, and amino(C 1 –C 6 )alkyl-;

R 3 is H, hydroxy, (C 1 –C 6 )alkoxy, —SOR 16 , —SO 2 R 17 , —C(O)OR 17 , —C(O)NR 18 R 19 , —(C 1 –C 6 )alkyl-C(O)NR 18 R 19 , (C 1 –C 6 )alkyl, halogen, fluoro(C 1 –C 6 )alkyl-, difluoro(C 1 –C 6 )alkyl-, trifluoro(C 1 –C 6 )alkyl-, (C 3 –C 6 )cycloalkyl, (C 3 –C 6 )-cycloalkyl-(C 1 –C 6 )alkyl-, (C 2 –C 6 )alkenyl, aryl(C 1 –C 6 )alkyl-, aryl(C 2 –C 6 )alkenyl-, heteroaryl(C 1 –C 6 )alkyl-, heteroaryl(C 2 –C 6 )alkenyl-, hydroxy(C 1 –C 6 )-alkyl-, —NR 22 R 23 , NR 22 R 23 —(C 1 –C 6 )alkyl-, aryl, thio(C 1 –C 6 )alkyl-, (C 1 –C 6 )alkyl-thio(C 1 –C 6 )alkyl-, (C 1 –C 6 )alkoxy(C 1 –C 6 )alkyl-, NR 18 R 19 —C(O)—(C 1 –C 6 )alkyl- or (C 3 –C 6 )cycloalkyl-(C 1 –C 6 )alkyl-;

Het is a mono- or bi-cyclic heteroaryl group of 5 to 10 atoms comprised of 1 to 9 carbon atoms and 1 to 4 heteroatoms independently selected from the group consisting of N, O and S, wherein a ring nitrogen can form an N-oxide or a quaternary group with a C 1 –C 4 alkyl group, wherein Het is attached to B by a carbon atom ring member, and wherein the Het group is substituted by W;

W is 1 to 4 substituents independently selected from the group consisting of H, (C 1 –C 6 )alkyl, fluoro(C 1 –C 6 )alkyl-, difluoro(C 1 –C 6 )alkyl-, trifluoro(C 1 –C 6 )alkyl-, (C 3 –C 6 )cycloalkyl, hydroxy(C 1 –C 6 )alkyl-, dihydroxy(C 1 –C 6 )alkyl-, NR 25 R 26 (C 1 –C 6 )alkyl-, thio(C 1 –C 6 )alkyl-, —OH, (C 1 –C 6 )alkoxy, halogen, —NR 4 R 5 , —C(O)OR 17 , —C(O)R 16 , (C 1 –C 6 )alkylthio-, R 21 -aryl, R 21 -aryl(C 1 –C 6 )alkyl-, aryl wherein adjacent carbons form a ring comprising a methylenedioxy group, and R 21 -heteroaryl;

R 4 and R 5 are independently selected from the group consisting of H, (C 1 –C 6 )alkyl, phenyl, benzyl and (C 3 –C 6 )cycloalkyl, or R 4 and R 5 taken together are —(CH 2 ) 4 —, —(CH 2 ) 5 — or —(CH 2 ) 2 NR 7 —(CH 2 ) 2 — and form a ring with the nitrogen to which they are attached;

R 6 is H, (C 1 –C 6 )alkyl or phenyl;

R 7 is H, (C 1 –C 6 )alkyl, —C(O)—R 16 , —C(O)OR 17 or —SO 2 R 17 ;

R 8 , R 10 and R 11 are independently selected from the group consisting of R 1 and —OR 1 , provided that when the optional double bond is present, R 10 is absent;

R 9 is H, OH or (C 1 –C 6 )alkoxy;

B is —(CH 2 ) n3 —, cis or trans —(CH 2 ) n4 CR 12 ═CR 12a (CH 2 ) n5 or —(CH 2 ) n4 C≡C(CH 2 ) n5 —, wherein n 3 is 0–5, n 4 and n 5 are independently 0–2, and R 12 and R 12a are independently selected from the group consisting of H, (C 1 –C 6 )alkyl and halogen;

X is —O— or —NR 6 — when the dotted line represents a single bond, or X is —OH or —NHR 20 when the bond is absent;

Y is ═O, ═S, (H, H), (H, OH) or (H, (C 1 –C 6 )alkoxy) when the dotted line represents a single bond, or when the bond is absent, Y is ═O, (H, H), (H, OH), (H, SH) or (H, (C 1 –C 6 )alkoxy);

each R 13 is independently selected from H, (C 1 –C 6 )alkyl, (C 3 –C 8 )cycloalkyl, —(CH 2 ) n6 NHC(O)OR 16b , —(CH 2 ) n6 NHC(O)R 16b , —(CH 2 ) n6 NHC(O)NR 4 R 5 , —(CH 2 ) n6 NHSO 2 R 16 , —(CH 2 ) n6 NHSO 2 NR 4 R 5 , and —(CH 2 ) n6 C(O)NR 28 R 29 where n 6 is 0–4, haloalkyl, and halogen;

each R 14 is independently selected from H, (C 1 –C 6 )alkyl, —OH, (C 1 –C 6 )alkoxy, R 27 -aryl(C 1 –C 6 )alkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, —(CH 2 ) n6 NHC(O)OR 16b , —(CH 2 ) n6 NHC(O)R 16b , —(CH 2 ) n6 NHC(O)NR 4 R 5 , —(CH 2 ) n6 NHSO 2 R 16 , —(CH 2 ) n6 NHSO 2 NR 4 R 5 , and —(CH 2 ) n6 C(O)NR 28 R 29 where n 6 is 0–4, halogen and haloalkyl; or

R 13 and R 14 taken together form a spirocyclic or a heterospirocyclic ring of 3–6 atoms;

wherein at least one of R 13 or R 14 is selected from the group consisting of —(CH 2 ) n6 NHC(O)OR 16b , —(CH 2 ) n6 NHC(O)R 16b , —(CH 2 ) n6 NHC(O)NR 4 R 5 , —(CH 2 ) n6 NHSO 2 R 16 , —(CH 2 ) n6 NHSO 2 NR 4 R 5 , and —(CH 2 ) n6 C(O)NR 28 R 29 where n 6 is 0–4;

R 15 is absent when the dotted line represents a single bond and is H, (C 1 –C 6 )alkyl, —NR 18 R 19 , or —OR 17 when the bond is absent;

R 16 is independently selected from the group consisting of (C 1 –C 6 )alkyl, phenyl and benzyl;

R 16b is H, alkoxy, (C 1 –C 6 )alkyl, (C 1 –C 6 )alkoxy(C 1 –C 6 )alkyl-, R 22 —O—C(O)—(C 1 –C 6 )alkyl-, (C 3 –C 6 )cycloalkyl, R 21 -aryl, R 21 -aryl(C 1 –C 6 )alkyl, haloalkyl, alkenyl, halosubstituted alkenyl, alkynyl, halosubstituted alkynyl, R 21 -heteroaryl, R 21 —(C 1 –C 6 )alkyl heteroaryl, R 21 —(C 1 –C 6 )alkyl heterocycloalkyl, R 28 R 29 N—(C 1 –C 6 )alkyl, R 28 R 29 N—(CO)—(C 1 –C 6 )alkyl, R 28 R 29 N—(CO)O—(C 1 –C 6 )alkyl, R 28 O(CO)N(R 29 )—(C 1 –C 6 )alkyl, R 28 S(O) 2 N(R 29 )—(C 1 –C 6 )alkyl, R 28 R 29 N—(CO)—N(R 29 )—(C 1 –C 6 )alkyl, R 28 R 29 N—S(O)2N(R 29 )—(C 1 –C 6 )alkyl, R 28 —(CO)N(R 29 )—(C 1 –C 6 )alkyl, R 28 R 29 N—S(O) 2 —(C 1 –C 6 )alkyl, HOS(O) 2 —(C 1 –C 6 )alkyl, (OH) 2 P(O) 2 —(C 1 –C 6 )alkyl, R 28 —S—(C 1 –C 6 )alkyl, R 28 —S(O) 2 —(C 1 –C 6 )alkyl or hydroxy(C 1 –C 6 )alkyl);

R 17 , R 18 and R 19 are independently selected from the group consisting of H, (C 1 –C 6 )alkyl, phenyl, and benzyl;

R 20 is H, (C 1 –C 6 )alkyl, phenyl, benzyl, —C(O)R 6 or —SO 2 R 6 ;

R 21 is 1 to 3 substituents independently selected from the group consisting of H, —CN, —CF 3 , —OCF 3 , halogen, —NO 2 , (C 1 –C 6 )alkyl, —OH, (C 1 –C 6 )alkoxy, (C 1 –C 6 )-alkylamino-, di-((C 1 –C 6 )alkyl)amino-, NR 25 R 26 -(C 1 –C 6 )alkyl-, hydroxy-(C 1 –C 6 )alkyl-, —C(O)OR 17 , —COR 17 , —NHCOR 16 , —NHSO 2 R 16 , —NHSO 2 CH 2 CF 3 , —C(O)NR 25 R 26 , —NR 25 —C(O)—NR 25 R 26 , —S(O)R 13 , —S(O) 2 R 13 and —SR 13 ;

R 22 is H or (C 1 –C 6 )alkyl;

R 23 is H, (C 1 –C 6 )alkyl, —C(O)R 24 , —SO 2 R 24 , —CONHR 24 or —SO 2 NHR 24 ;

R 24 is (C 1 –C 6 )alkyl, hydroxy (C 1 –C 6 )alkyl or NR 25 R 26 —((C 1 –C 6 )alkyl)-;

R 25 and R 26 are independently selected from the group consisting of H and (C 1 –C 6 )alkyl;

R 27 is 1, 2 or 3 substituents selected from the group consisting of H, (C 1 –C 6 )alkyl, (C 3 –C 6 )cycloalkyl, (C 1 –C 6 )alkoxy, halogen and —OH; and

R 28 and R 29 are independently selected from the group consisting of H, (C 1 –C 6 )alkyl, (C 1 –C 6 )alkoxy, R 27 -aryl(C 1 –C 6 )alkyl, heteroaryl, heteroarylalkyl, hydroxy(C 1 –C 6 )alkyl, (C 1 –C 6 )alkoxy(C 1 –C 6 )alkyl, heterocyclyl, heterocyclylalkyl, and haloalkyl; or

R 28 and R 29 taken together form a spirocyclic or a heterospirocyclic ring of 3–6 atoms.

Thrombin receptor antagonist compounds of the present invention can have anti-thrombotic, anti-platelet aggregation, antiatherosclerotic, antirestenotic and/or anti-coagulant activity. Thrombosis-related diseases treated by the compounds of this invention include thrombosis, atherosclerosis, restenosis, hypertension, angina pectoris, arrhythmia, heart failure, myocardial infarction, glomerulonephritis, thrombotic and thromboembolytic stroke, peripheral vascular diseases, other cardiovascular diseases, cerebral ischemia, inflammatory disorders and cancer, as well as other disorders in which thrombin and its receptor play a pathological role.

Certain embodiments of this invention also relate to a method of using at least one compound of Formula I in combination with one or more additional cardiovasular agents for the treatment of thrombosis, platelet aggregation, coagulation, cancer, inflammatory diseases or respiratory diseases, comprising administering a combination of at least one compound of formula I and at least one additional cardiovascular agent to a mammal in need of such treatment. In particular, the present invention relates to a method of using said combination in the treatment of thrombosis, atherosclerosis, restenosis, hypertension, angina pectoris, arrhythmia, heart failure, myocardial infarction, glomerulonephritis, thrombotic stroke, thromboembolic stroke, peripheral vascular diseases, cerebral ischemia, cancer, rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, diabetes, osteoporosis, renal ischemia, cerebral stroke, cerebral ischemia, nephritis, inflammatory disorders of the lungs and gastrointestinal tract, reversible airway obstruction, chronic asthma or bronchitis. It is contemplated that a combination of this invention may be useful in treating more than one of the diseases listed.

Some embodiments of the invention relate to a pharmaceutical composition comprising a therapeutically effective amount of a combination of at least one compound of formula I and at least one additional cardiovascular agent in a pharmaceutically acceptable carrier.

Some embodiments of the invention relate to the use of a thrombin receptor antagonist disclosed in any of U.S. Pat. Nos. 6,063,847, 6,326,380, U.S. Ser. Nos. 09/880,222 and 10/271,715, all of which are incorporated herein by reference, in combination with one or more additional cardiovascular agents, for the treatment of thrombosis, platelet aggregation, coagulation, cancer, inflammatory diseases or respiratory diseases. In particular, the present invention relates to a method of using said combination in the treatment of thrombosis, atherosclerosis, restenosis, hypertension, angina pectoris, arrhythmia, heart failure, myocardial infarction, glomerulonephritis, thrombotic stroke, thromboembolic stroke, peripheral vascular diseases, cerebral ischemia, cancer, rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, diabetes, osteoporosis, renal ischemia, cerebral stroke, cerebral ischemia, nephritis, inflammatory disorders of the lungs and gastrointestinal tract, reversible airway obstruction, chronic asthma or bronchitis.

It is further contemplated that the combination of the invention can be provided as a kit comprising in a single package at least one compound of formula I in a pharmaceutical composition, and at least one separate pharmaceutical composition comprising a cardiovascular agent.

For compounds of Formula I, preferred definitions of the variables are as follows:

The variable n is preferably 0–2, and more preferably 0. The optional double bond is preferably absent (i.e., the bond is a single bond).

Q is preferably:

with the six-membered Q ring being more preferred. R 13 is preferably H or —CH 3 . R 14 is preferably H or —CH 3 . For the five-membered Q ring, preferably no more than two R 13 and R 14 substituents are other than hydrogen. For the six-membered Q ring, preferably no more than four R 13 and R 14 substituents are other than hydrogen, more preferably no more than two R 13 and R 14 substituents are other than hydrogen.

Especially preferred Q rings are:

preferably shown as

respectively.

In the preferred Q rings above, R is preferably —(CH 2 ) n6 NHC(O)OR 16b , —(CH 2 ) n6 NHC(O)R 16b , —(CH 2 ) n6 NHC(O)NR 4 R 5 , —(CH 2 ) n6 NHSO 2 R 16 or —(CH 2 ) n6 NHSO 2 NR 4 R 5 wherein n 6 is 0–2, and R 16b , R 16 and R 4 are (C 1 –C 6 )alkyl and R 5 is H. More preferred are compounds of Formula I wherein R is —NHC(O)OR 16b , —NHC(O)R 16b , —NHC(O)NR 4 R 5 , —NHSO 2 R 16 or —NHSO 2 NR 4 R 5 wherein R 16b , R 16 and R 4 are (C 1 –C 6 )alkyl and R 5 is H. Even more preferred are compounds of Formula I wherein R is —NHC(O)OR 16b , —NHC(O)R 16b or —NHC(O)NR 4 R 5 , wherein R 16b and R 4 are (C 1 –C 6 )alkyl and R 5 is H.

R 1 and R 2 are preferably independently selected from the group consisting of H and (C 1 –C 6 )alkyl; more preferably, R 1 is (C 1 –C 6 )alkyl and R 2 is H; especially preferred are compounds wherein R 1 is —CH 3 and R 2 is H.

R 3 is preferably H, —OH, (C 1 –C 6 )alkyl, (C 1 –C 6 )alkoxy, halogen, (C 3 –C 6 )cycloalkyl, —C(O)OR 17 or —NR 22 R 23 ; more preferably, R 3 is H or (C 1 –C 6 )alkyl.

Het is preferably pyridyl attached to B by a carbon ring member, and is preferably substituted by 1 or 2 substituents selected from W, more preferably 1 substituent. W is preferably R 21 -aryl or R 21 -heteroaryl. Aryl is preferably phenyl. Heteroaryl is preferably pyridyl. R 21 is preferably H, halogen or —CN, or —CF 3 , especially F, —CN or —CF 3 .

R 8 , R 10 and R 11 are each independently preferably H or (C 1 –C 6 )alkyl, more preferably H or —CH 3 ; especially preferred are compounds of Formula I wherein R 8 , R 10 and R 11 are each H.

R 9 is preferably H, OH or (C 1 –C 6 )alkoxy; more preferably, R 9 is H.

B is preferably cis or trans —(CH 2 ) n4 CR 12 ═CR 12a (CH 2 ) n5 — wherein n 4 , n 5 , R 12 and R 12a are as defined above; more preferably, R 12 and R 12a are each H, and n 4 and n 5 are each zero. Particularly preferred are compounds wherein B is trans-alkenyl, especially —CH═CH—.

One group of preferred compounds is that wherein the optional single bond is present, X is —O—, Y is ═O, and R 15 is absent. Another preferred group of compounds is that wherein the optional single bond is absent, X is —OH, Y is (H, OH) and R 15 is H. Compounds wherein the optional single bond is present, X is —O—, Y is ═O, and R 15 is absent are more preferred.

Especially preferred are compounds of Formula I wherein R is —NHC(O)OR 16b wherein R 16b is (C 1 –C 6 )alkyl. R 16b is preferably methyl or ethyl. Also preferred are compounds wherein the R group is attached to the C-7 position of the Q ring, as shown in Formula IA below.

A preferred embodiment of the invention is a compound of Formula IA:

wherein R 1 , R 2 , R 3 , R 8 , R 10 , R 11 , B, and Het are defined as preferred above. At least one of ring carbon atoms 5–8 is preferably substituted with —(CH 2 ) n6 NHC(O)OR 16b , —(CH 2 ) n6 NHCOR 16b , —(CH 2 ) n6 NHCONR 4 R 5 , —(CH 2 ) n6 NHSO 2 R 16 or —(CH 2 ) n6 NHSO 2 NR 4 R 5 wherein n 6 is 0–2, and R 16b , R 16 and R 4 are (C 1 –C 6 )alkyl and R 5 is H.

A more preferred embodiment of the invention is a compound of Formula IB:

wherein Het is pyridyl substituted by an R 21 -aryl group, preferably an R 21 -phenyl group wherein R 21 is preferably F or —CF 3 .

Especially preferred are compounds of Formula IA or 1 B wherein at least one of ring carbon atoms 5–8 is —NHC(O)OR 16b wherein R 16b is (C 1 –C 6 )alkyl. R 16b is preferably methyl or ethyl.

As used above, and throughout the specification, the following terms, unless otherwise indicated, shall be understood to have the following meanings:

“Subject” includes both mammals and non-mammalian animals.

“Mammal” includes humans and other mammalian animals.

The term “optionally substituted” means optional substitution with the specified groups, radicals or moieties. It should be noted that any atom with unsatisfied valences in the text, schemes, examples and tables herein is assumed to have the hydrogen atom(s) to satisfy the valences.

The following definitions apply regardless of whether a term is used by itself or in combination with other terms, unless otherwise indicated. Therefore, the definition of “alkyl” applies to “alkyl” as well as the “alkyl” portions of “hydroxyalkyl”, “haloalkyl”, “alkoxy”, etc.

As used herein, the term “alkyl” means an aliphatic hydrocarbon group that can be straight or branched and comprises 1 to about 20 carbon atoms in the chain. Preferred alkyl groups comprise 1 to about 12 carbon atoms in the chain. More preferred alkyl groups comprise 1 to about 6 carbon atoms in the chain. “Branched” means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain. The alkyl can be substituted by one or more substituents independently selected from the group consisting of halo, aryl, cycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, —NH(alkyl), —NH(cycloalkyl), —N(alkyl) 2 (which alkyls can be the same or different), carboxy and —C(O)O-alkyl. Non-limiting examples of suitable alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, heptyl, nonyl, decyl, fluoromethyl, trifluoromethyl and cyclopropylmethyl.

“Alkenyl” means an aliphatic hydrocarbon group (straight or branched carbon chain) comprising one or more double bonds in the chain and which can be conjugated or unconjugated. Useful alkenyl groups can comprise 2 to about 15 carbon atoms in the chain, preferably 2 to about 12 carbon atoms in the chain, and more preferably 2 to about 6 carbon atoms in the chain. The alkenyl group can be substituted by one or more substituents independently selected from the group consisting of halo, alkyl, aryl, cycloalkyl, cyano and alkoxy. Non-limiting examples of suitable alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut-enyl and n-pentenyl.

Where an alkyl or alkenyl chain joins two other variables and is therefore bivalent, the terms alkylene and alkenylene, respectively, are used.

“Alkoxy” means an alkyl-O— group in which the alkyl group is as previously described. Useful alkoxy groups can comprise 1 to about 12 carbon atoms, preferably 1 to about 6 carbon atoms. Non-limiting examples of suitable alkoxy groups include methoxy, ethoxy and isopropoxy. The alkyl group of the alkoxy is linked to an adjacent moiety through the ether oxygen.

“Alkynyl” means an aliphatic hydrocarbon group comprising at least one carbon-carbon triple bond and which may be straight or branched and comprising about 2 to about 15 carbon atoms in the chain. Preferred alkynyl groups have about 2 to about 12 carbon atoms in the chain; and more preferably about 2 to about 4 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkynyl chain. Non-limiting examples of suitable alkynyl groups include ethynyl, propynyl, 2-butynyl, 3-methylbutynyl, n-pentynyl, and decynyl. The alkynyl group may be substituted by one or more substituents which may be the same or different, each substituent being independently selected from the group consisting of alkyl, aryl and cycloalkyl.

“Aryl” means an aromatic monocyclic or multicyclic ring system comprising about 5 to about 14 carbon atoms, preferably about 6 to about 10 carbon atoms. The aryl can be substituted with one or more substituents, as defined above, which may be the same or different. Non-limiting examples of suitable aryl groups include phenyl, naphthyl, indenyl, tetrahydronaphthyl and indanyl. “Arylene” means a bivalent phenyl group, including ortho, meta and para-substitution.

The term “Boc” refers to N-tert-butoxycarbonyl.

“Cycloalkyl” means a non-aromatic mono- or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 5 to about 10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7 ring atoms. The cycloalkyl can be substituted with one or more substituents, as defined above, which may be the same or different. Non-limiting examples of suitable monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. Non-limiting examples of suitable multicyclic cycloalkyls include 1-decalinyl, norbornyl, adamantyl and the like. “Cycloalkylene” refers to a corresponding bivalent ring, wherein the points of attachment to other groups include all positional isomers.

“Dihydroxy(C 1 –C 6 )alkyl” refers to an alkyl chain substituted by two hydroxy groups on two different carbon atoms.

“Fluoroalkyl”, “difluoroalkyl” and “trifluoroalkyl” mean alkyl chains wherein the terminal carbon is substituted by 1, 2 or 3 fluoroatoms, respectively, e.g., —CF 3 , —CH 2 CF 3 , —CH 2 CHF 2 or —CH 2 CH 2 F.

“Halogen” or “halo” refers to fluorine, chlorine, bromine or iodine radicals. Preferred are fluoro, chloro or bromo, and more preferred are fluoro and chloro.

“Heteroaryl” means a single ring, bicyclic or benzofused heteroaryl group of 5 to 14 ring atoms, preferably about 5 to 10 ring atoms, comprised of 1 to 13 carbon atoms and 1 to 4 heteroatoms independently selected from the group consisting of N, O and S, provided that the rings do not include adjacent oxygen and/or sulfur atoms. N-oxides of the ring nitrogens are also included, as well as compounds wherein a ring nitrogen is substituted by a (C 1 –C 4 )alkyl group to form a quaternary amine. Examples of single-ring heteroaryl groups are pyridyl, oxazolyl, isoxazolyl, oxadiazolyl, furanyl, pyrrolyl, thienyl, imidazolyl, pyrazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyrazinyl, pyrimidyl, pyridazinyl and triazolyl. Examples of bicyclic heteroaryl groups are naphthyridyl (e.g., 1, 5 or 1, 7), imidazopyridyl, pyrido[2,3]imidazolyl, pyridopyrimidinyl and 7-azaindolyl. Examples of benzofused heteroaryl groups are indolyl, quinolyl, isoquinolyl, phthalazinyl, benzothienyl (i.e., thionaphthenyl), benzimidazolyl, benzofuranyl, benzoxazolyl and benzofurazanyl. All positional isomers are contemplated, e.g., 2-pyridyl, 3-pyridyl and 4-pyridyl.

The term “Het” is exemplified by the single ring, bicyclic and benzofused heteroaryl groups as defined immediately above. Het groups are joined to group B by a carbon ring member, e.g., Het is 2-pyridyl, 3-pyridyl or 2-quinolyl. The Het ring can be substituted on any available ring carbon by a group W; 1 to 4 W substituents can be present on a Het ring.

“Heterocycloalkyl” means a 4 to 6 membered saturated ring containing 3 to 5 carbon atoms and 1 or 2 heteroatoms selected from the group consisting of N, S and O, provided that the heteroatoms are not adjacent. Examples of heterocycloalkyl rings are pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl and tetrahydrothiopyranyl.

The term “heterospirocyclic” refers to a spirocyclic structure containing 3 to 5 carbon atoms and 1 or 2 heteroatoms selected from the group consisting of N, S and O, provided that the heteroatoms are not adjacent.

The term “optional single bond” refers to the bond shown by the dotted line between X and the carbon to which Y and R 15 are attached in the structure of Formula I. The optional double bond represented by means that at least a single bond must be present, but that a double bond can be present; when the double bond is present, R 10 is absent.

When R 4 and R 5 join to form a ring with the nitrogen to which they are attached, the rings formed are 1-pyrrolidinyl, 1-piperidinyl and 1-piperazinyl, wherein the piperazinyl ring may also be substituted at the 4-position nitrogen by a group R 7 .

The above statements, wherein, for example, R 4 and R 5 are said to be independently selected from a group of substituents, means that R 4 and R 5 are independently selected when attached to the same nitrogen, but also that where an R 4 or R 5 variable occurs more than once in a molecule, those occurrences are independently selected. Similarly, each occurrence of R 13 or R 14 is independent of any other R 13 or R 14 in the same Q ring. Those skilled in the art will recognize that the size and nature of the substituent(s) will affect the number of substituents which can be present.

Compounds of the invention have at least one asymmetrical carbon atom and therefore all isomers, including enantiomers, stereoisomers, rotamers, tautomers and racemates of the compounds of Formula (I) (where they exist) are contemplated as being part of this invention. The invention includes d and I isomers in both pure form and in admixture, including racemic mixtures. Isomers can be prepared using conventional techniques, either by reacting optically pure or optically enriched starting materials or by separating isomers of a compound of Formula I. Isomers may also include geometric isomers, e.g., when a double bond is present. Polymorphous forms of the compounds of Formula (I), whether crystalline or amorphous, also are contemplated as being part of this invention.

Those skilled in the art will appreciate that for some of the compounds of Formula I, one isomer will show greater pharmacological activity than other isomers.

Typical preferred compounds of the present invention have the following stereochemistry:

with compounds having that absolute stereochemistry being more preferred.

Those skilled in the art will appreciate that for some compounds of Formula I, one isomer will show greater pharmacological activity than other isomers.

Compounds of the invention with a basic group can form pharmaceutically acceptable salts with organic and inorganic acids. Examples of suitable acids for salt formation are hydrochloric, sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic, maleic, methanesulfonic and other mineral and carboxylic acids well known to those in the art. Preferred embodiments include bisulfate salts. The salt is prepared by contacting the free base form with a sufficient amount of the desired acid to produce a salt. The free base form may be regenerated by treating the salt with a suitable dilute aqueous base solution such as dilute aqueous sodium bicarbonate. The free base form differs from its respective salt form somewhat in certain physical properties, such as solubility in polar solvents, but the salt is otherwise equivalent to its respective free base forms for purposes of the invention. Compounds of the invention can also form pharmaceutically acceptable solvates, including hydrates.

Certain compounds of the invention are acidic (e.g., those compounds which possess a carboxyl group). These compounds form pharmaceutically acceptable salts with inorganic and organic bases. Examples of such salts are the sodium, potassium, calcium, aluminum, lithium, gold and silver salts. Also included are salts formed with pharmaceutically acceptable amines such as ammonia, alkyl amines, hydroxyalkylamines, N-methylglucamine and the like.

“Solvate” means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. “Solvate” encompasses both solution-phase and isolatable solvates. Non-limiting examples of suitable solvates include ethanolates, methanolates, and the like. “Hydrate” is a solvate wherein the solvent molecule is H 2 O.

Compounds of the present invention in which n 6 is 0 can be prepared by processes known in the art, for example by the processes described in U.S. Pat. No. 6,063,847, incorporated herein by reference, and by the processes exemplified below.

Compounds of the present invention in which n 6 is 1 or 2 are generally prepared by processes in accordance with the following general scheme:

Ketone 4 is subjected to a Wittig reaction to provide vinyl ether 5 which is hydrolyzed under acidic condition to provide aldehyde 6. The aldehyde is reduced to the alcohol 7 and converted to azide 8 via its mesylate. Reduction of the azido group with Me 3 P provides the amine 9 which is treated with different electrophiles to provide diverse analogs.

Following are examples of preparing compounds of Formula I. In the examples, the following abbreviations are used: Et (ethyl); Me (methyl); Pr (propyl); Ac (acetyl); rt (room temperature); PTLC (preparative thin layer chromatography); THF (tetrahydrofuran); TBAF (tetra-n-butylammonium fluoride); Tips (triisopropylsilyl); and Tf (trifluoromethanesulfonyl).

Compound 1, described in U.S. Pat. No. 6,063,847, (1.95 g, 4.2 mmol) was dissolved in EtOH (40 ml) and CH 2 Cl 2 (10 ml). Then, NH 3 (g) was bubbled into the solution for 5 min. The reaction mixture was C(O)Oled to 0° C., and Ti(OiPr) 4 (1.89 ml, 6.3 mmol) was added. After stirring at 0° C. for 1 h, 1M TiCl 4 (6.3 ml, 6.3 mmol) was added. The reaction mixture was stirred at rt for an additional 45 min and concentrated to dryness under reduced pressure. The residue was dissolved in CH 3 OH (10 ml) and NaBH 3 CN (510 mg, 8 mmol) was added. The resulting suspension was stirred overnight at rt. The reaction mixture was poured to 1 N NaOH (100 ml) and extracted with EtOAc (3×100 ml). The organic layer was combined and dried with Na 2 SO 4 . Removal of solvent afforded product 2 (1.2 g, 62%). Further separation by PTLC (5% 2 M NH 3 in CH 3 OH/CH 2 Cl 2 ) afforded β-2 (spot 1) and α-2 (spot 2) in a 1:2 ratio. β-2: 1 HNMR (CDCl 3 ): δ 0.81–1.15 (m, 2H), 1.11–1.38 (m, 4H), 1.42 (d, J=6 Hz, 3H), 1.82–2.01 (m, 3H), 2.37 (m, 2H), 2.45 (br m, 1H), 2.65 (m, 1H), 2.81 (m, 1H), 4.75 (m, 1H), 6.61 (m, 2H), 7.26 (m, 2H), 7.75–7.85 (m, 4H), 8.77 (d, J=1.6 Hz, 1H). α-2: 1 HNMR (CDCl 3 ): δ 0.95 (m, 1H), 1.10–1.40 (m, 5H), 1.41 (d, J=6 Hz, 3H), 1.52–1.65 (m, 2H), 1.75 (m, 1H), 1.84–2.0 (m, 2H), 2.37 (m, 1H), 2.45 (m, 1H), 2.65 (m, 1H), 3.42 (br s, 1H), 4.70 (m, 1H), 6.61 (m, 2H), 7.26 (m, 2H), 7.75–7.85 (m, 4H), 8.77 (d, J=1.6 Hz, 1H).

Compound α-2 (110 mg), ethyl chloroformate (0.4 ml) and Et 3 N (0.5 ml) in CH 2 Cl 2 (6 ml) were stirred for 2 h. The reaction mixture was directly separated by PTLC (EtOAc/hexane, 1:1) to afford the title compound (100 mg, 79%). MS m/z 543 (M+1). HRMS Calcd for C 30 H 34 N 2 O 4 F 3 (M+1): 543.2471, found 543.2467.

N-Methyl Compound for Comparison

Compound 1 (646 mg, 1.38 mmol) was dissolved in 2.0 M CH 3 NH 2 in CH 3 OH (15 ml, 30 mmol) and stirred at rt for 5 min followed by addition of NaCNBH 3 (173 mg, 2.75 mmol). The reaction mixture was stirred at rt overnight and concentrated to dryness under reduced pressure. Removal of the solvent followed by PTLC separation (7% 1 M NH 3 in CH 3 OH/CH 2 Cl 2 ) afforded β-4 (spot 1, 76 mg, 11%) and α-4 (spot 2, 100 mg, 15%). β-4: 1 HNMR (CDCl 3 ): δ 1.15–1.24 (m, 5H), 1.42 (d, J=6 Hz, 3H), 1.42–1.61 (m, 2H), 1.71–1.95 (m, 4H), 2.21 (m, 1H), 2.38 (s, 3H), 2.45 (m, 1H), 2.71 (m, 1H), 2.84 (m, 1H), 4.75 (m, 1H), 6.51–6.63 (m, 2H), 7.26 (m, 2H), 7.75–7.85 (m, 4H), 8.77 (d, J=2.0 Hz, 1H). α-4: 1 HNMR (CDCl 3 ): δ 0.95 (m, 2H), 1.10–1.40 (m, 5H), 1.41 (d, J=6 Hz, 3H), 1.82–1.95 (m, 5H), 2.38 (m, 2H), 2.42 (s, 3H), 2.65 (m, 1H), 4.79 (m, 1H), 6.51–6.63 (m, 2H), 7.26 (m, 2H), 7.75–7.85 (m, 4H), 8.77 (d, J=2.0 Hz, 1H).

Compound α-4 (50 mg), ethyl chloroformate (0.15 ml) and Et 3 N (0.3 ml) in CH 2 Cl 2 (5 ml) were stirred for 2 h. The reaction mixture was directly separated by PTLC (EtOAc/hexane, 1:1) to afford the title compound (48 mg, 84%). MS m/z 557 (M+1). HRMS Calcd for C 31 H 36 N 2 O 4 F 3 (M+1): 557.2627, found 557.2620.

Phosphonate 7, described in U.S. Pat. No. 6,063,847, (3.27 g, 8.1 mmol) was dissolved in THF (12 ml) and C(O)Oled to 0° C., followed by addition of 2.5 M n-BuLi (3.2 ml, 8.1 mmol). The reaction mixture was stirred at 0° C. for 10 min and warmed up to rt. A solution of aldehyde 6, described in U.S. Pat. No. 6,063,847, in THF (12 ml) was added to the reaction mixture. The reaction mixture was stirred for 30 min. Standard aqueous work-up, followed by column chromatography (30–50% EtOAc in hexane) afforded product 8. 1 HNMR (CDCl 3 ): δ 0.92–1.38 (m, 31H), 1.41 (d, J=6 Hz, 3H), 1.40–1.55 (m, 2H), 1.70–1.80 (m, 2H), 1.81–1.90 (m, 2H), 2.36 (m, 2H), 2.69 (m, 1H), 3.89 (m, 4H), 4.75 (m, 1H), 6.28–6.41 (m, 2H), 7.05–7.15 (m, 2H), 8.19 (brs, 1H).

Compound 8 (2.64 g, 4.8 mmol) was dissolved in THF (48 ml). The reaction mixture was C(O)Oled to 0° C. followed by addition of 1 M TBAF (4.8 ml). The reaction mixture was stirred for 5 min followed by standard aqueous work-up. Column chromatography (50% EtOAc/hexane) afforded product 9 (1.9 g, 100%). 1 HNMR (CDCl 3 ):δ 1.15–1.55 (m, 6H), 1.41 (d, J=6 Hz, 3H), 1.70–1.82 (m, 3H), 1.85–1.90 (m, 1H), 2.36 (m, 2H), 2.69 (m, 1H), 3.91 (m, 4H), 4.75 (m, 1H), 6.18–6.45 (m, 2H), 7.19 (br s, 2H), 8.19 (br s, 1H).

To a solution of compound 9 (250 mg, 0.65 mmol) in pyridine (5 ml) C(O)Oled to 0° C. was added Tf 2 O (295 μL, 2.1 mmol). The reaction mixture was stirred overnight at rt. Standard aqueous work-up followed by column chromatography afforded product 10 (270 mg, 80%). 1 HNMR (CDCl 3 ): δ 1.15–1.55 (m, 6H), 1.41 (d, J=6 Hz, 3H), 1.70–1.82 (m, 3H), 1.85–1.90 (m, 1H), 2.36 (m, 2H), 2.69 (m, 1H), 3.91 (m, 4H), 4.75 (m, 1H), 6.42–6.68 (m, 2H), 7.25 (m, 1H), 7.55 (m, 1H), 8.49 (d, J=2.8 Hz, 1H).

Compound 10 (560 mg, 1.1 mmol), 3-fluorophenyl boronic acid (180 mg, 1.3 mmol) and K 2 CO 3 (500 mg, 3.6 mmol) were mixed with toluene (4.4 ml), H 2 O (1.5 ml) and EtOH (0.7 ml) in a sealed tube. Under an atmosphere of N 2 , Pd(Ph 3 P) 4 (110 mg, 0.13 mmol) was added. The reaction mixture was heated at 100° C. for 2 h under N 2 . The reaction mixture was C(O)Oled down to rt, poured to EtOAc (30 ml) and washed with water (2×20 ml). The EtOAc solution was dried with NaHCO 3 and concentrated at reduced pressure to give a residue. Preparative TLC separation of the residue (50% EtOAc in hexane) afforded product 11 (445 mg, 89%). 1 HNMR (CDCl 3 ): δ 1.15–1.59 (m, 6H), 1.43 (d, J=6 Hz, 3H), 1.70–1.79 (m, 2H), 1.82 (m, 1H), 1.91 (m, 2H), 2.41 (m, 2H), 2.69 (m, 1H), 3.91 (m, 4H), 4.75 (m, 1H), 6.52–6.68 (m, 2H), 7.15 (m, 1H), 7.22 (m, 2H), 7.35 (m, 1H), 7.44 (m, 1H), 7.81 (m, 1H), 8.77 (d, J=1.2 Hz, 1H).

Compound 11 (445 mg, 0.96 mmol) was dissolved in a mixture of acetone (10 ml) and 1 N HCl (10 ml). The reaction mixture was heated at 50° C. for 1 h. Standard aqueous work-up followed by preparative TLC separation (50% EtOAc in hexane) afforded product 12(356 mg, 89%). 1 HNMR (CDCl 3 ): δ 1.21–1.45 (m, 2H), 1.47 (d, J=5.6 Hz, 3H), 1.58–1.65 (m, 2H), 2.15 (m, 1H), 2.18–2.28 (m, 2H), 2.35–2.51 (m, 5H), 2.71 (m, 1H), 4.79 (m, 1H), 6.52–6.68 (m, 2H), 7.15 (m, 1H), 7.22 (m, 2H), 7.35 (m, 1H), 7.44 (m, 1H), 7.81 (m, 1H), 8.77 (d, J=1.2 Hz, 1H).

Compound 12 (500 mg, 4.2 mmol) was dissolved in EtOH (40 ml) and CH 2 Cl 2 (15 ml) NH 3 (g) was bubbled into the solution for 5 min. The reaction mixture was C(O)Oled to 0° C. followed by addition of Ti(OiPr) 4 (1.89 ml, 6.3 mmol). After stirring at 0° C. for 1 h, 1M TiCl 4 (6.3 ml, 6.3 mmol) was added. The reaction mixture was stirred at rt for 45 min and concentrated to dryness under reduced pressure. The residue was dissolved in CH 3 OH (10 ml) and NaBH 3 CN (510 mg, 8 mmol) was added. The reaction mixture was stirred overnight at rt. The reaction mixture was poured to 1 N NaOH (100 ml) and extracted with EtOAc (3×100 ml). The organic layer was combined and dried with NaHCO 3 . Removal of solvent and separation by PTLC (5% 2 M NH 3 in CH 3 OH/CH 2 Cl 2 ) afforded β-13 (spot 1, 30 mg, 6%) and α-13 (spot 2, 98 mg, 20%). β-13: 1 HNMR (CDCl 3 ): δ 1.50–1.38 (m, 5H), 1.42 (d, J=6 Hz, 3H), 1.51–1.75 (m, 5H), 1.84 (m, 2H), 2.38 (m, 1H), 2.45 (m, 1H), 3.38 (br s, 1H), 4.78 (m, 1H), 6.59 (m, 2H), 7.15 (m, 1H), 7.26 (m, 2H), 7.36 (m, 1H), 7.42 (m, 1H), 7.82 (m, 1H), 8.77 (d, J=2 Hz, 1H). α-13: 1 HNMR (CDCl 3 ): δ 0.95 (m, 2H), 1.02–1.35 (m, 6H), 1.41 (d, J=6 Hz, 3H), 1.82–1.95 (m, 4H), 2.37 (m, 2H), 2.69 (m, 2H), 4.71 (m, 1H), 6.71 (m, 2H), 7.11 (m, 1H), 7.25 (m, 2H), 7.38 (m, 1H), 7.42 (m, 1H), 7.80 (m, 1H), 8.76 (d, J=1.6 Hz, 1H).

Compound α-13 (300 mg, 0.71 mmol) was dissolved in CH 2 Cl 2 (10 ml) followed by addition of Et 3 N (0.9 ml). The reaction mixture was C(O)Oled to 0° C. and ethyl chloroformate (0.5 ml) was added. The reaction mixture was stirred at rt for 1 h. The reaction mixture was directly separated by preparative TLC (EtOAc/hexane, 1:1) to give the title compound (14) (300 mg, 86%). MS m/z 493 (M+1). HRMS Calcd for C 29 H 34 N 2 O 4 F (M+1): 493.2503, found 493.2509.

N-Methyl Compound for Comparison

Compound 12 (130 mg, 0.31 mmol) was dissolved in 2.0 M CH 3 NH 2 in CH 3 OH (5 ml, 10 mmol). After stirring at rt for 5 min, NaCNBH 3 (40 mg, 0.62 mmol) was added. The reaction mixture was stirred at rt overnight. Removal of the solvent followed by PTLC separation (7% 1 M NH 3 in CH 3 OH/CH 2 Cl 2 ) afforded β-15 (spot 1, 20 mg, 15%) and α-15 (spot 2, 25 mg, 19%). β-15: 1 HNMR (CDCl 3 ): δ 1.15–1.25 (m, 5H), 1.42 (m, 3H), 1.42–1.61 (m, 2H), 1.75–1.90 (m, 3H), 2.25–2.45 (m, 2H), 2.41 (s, 3H), 2.70 (m, 1H), 2.85 (m, 1H), 4.75 (m, 1H), 6.51–6.61 (m, 2H), 7.11 (m, 1H), 7.23–7.27 (m, 2H), 7.35 (m, 1H), 7.45 (m, 1H), 7.80 (m, 1H), 8.76 (d, J=2.4 Hz, 1H). α-15: 1 HNMR (CDCl 3 ): δ 0.90 (m, 2H), 1.10–1.35 (m, 5H), 1.41 (d, J=5.6 Hz, 3H), 1.82–2.01 (m, 4H), 2.36 (m, 2H), 2.39 (s, 3H), 2.55–2.65 (br s, 1H), 2.71 (m, 1H), 4.79 (m, 1H), 6.51–6.63 (m, 2H), 7.08 (m, 1H), 7.26 (m, 2H), 7.34 (m 1H), 7.42 (m, 1H), 7.81 (m, 1H), 8.76 (d, J=2.0 Hz, 1H).

Compound α-15 (25 mg, 0.06 mmol) was dissolved in CH 2 Cl 2 (5 ml) followed by addition of Et 3 N (0.2 ml). The reaction mixture was C(O)Oled to 0° C. and ethyl chloroformate (0.1 m) was added. The reaction mixture was stirred at rt for 1 h. The reaction mixture was directly separated by preparative TLC (EtOAc/hexane, 1:1) to give the title compound (25 mg, 85%). MS m/z 507 (M+1). HRMS Calcd for C 30 H 36 N 2 O 4 F (M+1): 507.2659, found 507.2652.

Compound 18A (0.100 g, 0.238 mmol) was stirred with ethyl chloroformate (0.195 ml, 2.38 mmol) and Et 3 N (0.5 ml, 3.6 mmol) in CH 2 Cl 2 (10 ml) at 0° C. for 10 min and at rt for 1 h. The mixture was diluted with EtOAc (50 ml) and washed with NaHCO 3 (sat.). The organic layer was dried (MgSO 4 ) and concentrated in vacuo. Flash chromatography of the residue on a silica gel column with EtOAc-hexane (50-50) as eluent gave Example 69A (100 mg, 85%). MS (ESI) m/z 493 (MH + ). Compound 32B was similarly prepared from 18B. MS (ESI) m/z 493 (MH + ).

Using the procedure of Example 32, Step 1, starting with compound 19 (see U.S. Pat. No. 6,063,847), prepare: 20A (lower R f ), MS (FAB) m/z 421 (MH + ), and 20B (higher R f ), MS (FAB) m/z 421 (MH + ).

Using the procedure of Example 69, Step 2, Example 70A was prepared from compound 20A: MS (ESI) m/z 493 (MH + ). Using the procedure of Example 69, Step 2, Example 70B was prepared from compound 20B: MS (ESI) m/z 493 (MH + ).